{"title":"用于细胞外囊泡检测的功能分子集成纳米界面","authors":"Hirobumi Sunayama, Yuya Matsui, Toshifumi Takeuchi","doi":"10.1002/adsr.202400186","DOIUrl":null,"url":null,"abstract":"<p>Functional molecule-integrated polymer nanocavities for the detection of extracellular vesicles (EVs) are prepared via template polymerization and multistep post-modification. A multifunctional polymer bearing a tertiary amino group for interaction with template silica nanoparticles (NPs) and a polymerizable methacryl group is prepared and complexed with 500-nm silica NPs. The complex is monodispersedly immobilized on the carboxyl group and Br group introduced onto the Au-coated glass substrate. Additionally, surface-initiated atom transfer radical polymerization of 2-methacryloyloxyethyl phosphorylcholine is conducted to form a biocompatible polymer layer. After the silica NPs are removed, the disulfide bond is cleaved using tris(2-carboxyethyl)phosphine to remove the multifunctional polymer components, yielding a thiol group-exposed nanocavity. The degree of functionalization is controlled by changing the mixing ratio of the fluorescent and non-fluorescent dyes. A multifunctionalized nanocavity is prepared by introducing a fluorescent dye and nitrilotriacetic acid moiety, followed by His-tagged protein G and an antibody for the CD9 protein, yielding EV-sensing nano-interfaces. This fluorescent signaling interface responds to EVs in a concentration-dependent manner, with an estimated limit of detection of 0.1 fM. These responses are not observed when the reference antibody is introduced, confirming cavity-selective functionalization. The proposed method is useful for fabricating multifunctional nano-interfaces in the field of life sciences.</p>","PeriodicalId":100037,"journal":{"name":"Advanced Sensor Research","volume":"4 6","pages":""},"PeriodicalIF":3.5000,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/adsr.202400186","citationCount":"0","resultStr":"{\"title\":\"Functional Molecule-Integrated Nano-Interfaces for Detection of Extracellular Vesicles\",\"authors\":\"Hirobumi Sunayama, Yuya Matsui, Toshifumi Takeuchi\",\"doi\":\"10.1002/adsr.202400186\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Functional molecule-integrated polymer nanocavities for the detection of extracellular vesicles (EVs) are prepared via template polymerization and multistep post-modification. A multifunctional polymer bearing a tertiary amino group for interaction with template silica nanoparticles (NPs) and a polymerizable methacryl group is prepared and complexed with 500-nm silica NPs. The complex is monodispersedly immobilized on the carboxyl group and Br group introduced onto the Au-coated glass substrate. Additionally, surface-initiated atom transfer radical polymerization of 2-methacryloyloxyethyl phosphorylcholine is conducted to form a biocompatible polymer layer. After the silica NPs are removed, the disulfide bond is cleaved using tris(2-carboxyethyl)phosphine to remove the multifunctional polymer components, yielding a thiol group-exposed nanocavity. The degree of functionalization is controlled by changing the mixing ratio of the fluorescent and non-fluorescent dyes. A multifunctionalized nanocavity is prepared by introducing a fluorescent dye and nitrilotriacetic acid moiety, followed by His-tagged protein G and an antibody for the CD9 protein, yielding EV-sensing nano-interfaces. This fluorescent signaling interface responds to EVs in a concentration-dependent manner, with an estimated limit of detection of 0.1 fM. These responses are not observed when the reference antibody is introduced, confirming cavity-selective functionalization. The proposed method is useful for fabricating multifunctional nano-interfaces in the field of life sciences.</p>\",\"PeriodicalId\":100037,\"journal\":{\"name\":\"Advanced Sensor Research\",\"volume\":\"4 6\",\"pages\":\"\"},\"PeriodicalIF\":3.5000,\"publicationDate\":\"2025-04-21\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://onlinelibrary.wiley.com/doi/epdf/10.1002/adsr.202400186\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Advanced Sensor Research\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/adsr.202400186\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Advanced Sensor Research","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/adsr.202400186","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Functional Molecule-Integrated Nano-Interfaces for Detection of Extracellular Vesicles
Functional molecule-integrated polymer nanocavities for the detection of extracellular vesicles (EVs) are prepared via template polymerization and multistep post-modification. A multifunctional polymer bearing a tertiary amino group for interaction with template silica nanoparticles (NPs) and a polymerizable methacryl group is prepared and complexed with 500-nm silica NPs. The complex is monodispersedly immobilized on the carboxyl group and Br group introduced onto the Au-coated glass substrate. Additionally, surface-initiated atom transfer radical polymerization of 2-methacryloyloxyethyl phosphorylcholine is conducted to form a biocompatible polymer layer. After the silica NPs are removed, the disulfide bond is cleaved using tris(2-carboxyethyl)phosphine to remove the multifunctional polymer components, yielding a thiol group-exposed nanocavity. The degree of functionalization is controlled by changing the mixing ratio of the fluorescent and non-fluorescent dyes. A multifunctionalized nanocavity is prepared by introducing a fluorescent dye and nitrilotriacetic acid moiety, followed by His-tagged protein G and an antibody for the CD9 protein, yielding EV-sensing nano-interfaces. This fluorescent signaling interface responds to EVs in a concentration-dependent manner, with an estimated limit of detection of 0.1 fM. These responses are not observed when the reference antibody is introduced, confirming cavity-selective functionalization. The proposed method is useful for fabricating multifunctional nano-interfaces in the field of life sciences.